1.Cloning and expression of genotype B and C hepatitis B virus in eukaryotic cells
Xiaoguang LI ; Yuan HONG ; Qi WANG ; Jinqian ZHANG ; Jun CHENG
Chinese Journal of Infectious Diseases 2010;28(1):10-13
Objective To construct recombinant full length genotype B and C hepatitis B virus (HBV)and to examine HBV DNA replication and hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)expressions in Huh7 cells. Methods The full length genotype B and C HBV DNA were extracted and amplified from two HBV infected patients. The recombinant plasmids were constructed by inserting the amplified HBV fragments into the eukaryotic expression vector,pHY106,which were then transfected into Huh7 cells. The cells transfected with blank pHY106 vector were used as control. HBV DNA replication at 72 hours of transfection was detected by Southern blot. The HBV DNA levels in Huh7 cells at 24,48,72,96 and 120 hours of transfection were determined by real-time polymerase chain reaction(PCR).Meanwhile, the HBsAg and HBeAg expression levels in the supernatants at 24,48,72,96 and 120 hours were determined by enzyme linked immunosorbent assay(ELISA).Results The recombinant plasmids expressing genotype B or C HBV DNA were successfully constructed.The HBV replicative intermediates in HBV core particles,including rcDNA dsDNA and ssDNA,were detected by Southern blot.HBV DNA level could reach 8 lg copy/mL which was by real-time PCR. HBsAg and HBeAg levels determined by ELISA peaked at 72 hours after transfection and then declined gradually. Conclusions The recombinant plasmids inserted with genotype B or C HBV DNA are constructed successfully, which can express high levels of HBsAg and HBeAg in Huh7 cells. This system provides a platform for studying the pathogenesis of B and C genotype HBV, the interaction between HBV and host, as well as exploiting new drugs against HBV.
2.Mutual interaction between hepatitis C virus core protein and apolipoprotein A Ⅰ
Jing SHEN ; Jun CHENG ; Chaopin LI ; Xiaoquan LI ; Qi WANG ; Guoli LI ; Jinqian ZHANG
Chinese Journal of Microbiology and Immunology 2009;29(8):697-700
group pACT-apoA Ⅰ and pBIND-core was higher than that in the negative control.Conclusion HCV core protein and apoA Ⅰ can interact in vivo.This study provides new theory for the mechanism of chronicity hepatitis C in fatty liver.
3.Experiment study of three dimensional navigation assisted spinal surgery using structured light scanning
Bin FENG ; Guixing QIU ; Shugang LI ; Haojun ZHENG ; Pengcheng LI ; Siyi CAI ; Jinqian LIANG ; Lin SHENG
Chinese Journal of Orthopaedics 2011;31(5):530-534
Objective To introduce a new spinal surgery navigation system based on structured light scanning(structured light navigation system,SLNS),and to compare accuracy of pedicle screw placement using SLNS,CT based navigation system and free hand technique.Methods Thirty-two calf vertebral pedicles,40 and 32 were placed with pedicle screws using SLNS,CT navigation system,and free hand technique,respectively.The pedicle breakage ratio,transverse section angle (TSA),sagittal section angle (SSA),screw offset deviation of each screw were detected according to CT images.Results The pedicle breakage ratio of pedicle screws in SLNS group,CT navigation group and free hand group were 6%(2./32),5%(2/40),25%(8/32),respectively.The difference between each of the two navigation groups and free hand group were statistically significant regarding frequency of screw misplacement.The difference between the two navigation systems was not obvious.The average SSA error and TSA error in SLNS group were 2.58°±2.74° and 4.26°±5.20°.For CT navigation group,they were 2.95°±2.61° and 3.13°±-2.75°.There was no statistical difference between the two navigation methods for the SSA or TSA error.There was no statistical difference in offset deviation among SLNS group,CT navigation group and free hand group.Conclusion The SLNS is a new and practical navigation system,which has similar accuracy with CT navigation system.
4.Screening of proteins binding to hepatitis C virus E1 protein from human pancreas cDNA library
Xiaochun WANG ; Jinqian ZHANG ; Shuling WU ; Chenyu ZHANG ; Chaopin LI ; Guoli LI ; Qi WANG ; Jun CHENG
Chinese Journal of Infectious Diseases 2008;26(11):647-650
Objective To screen proteins from human pancreas cDNA library,which interact with hepatitis C virus(HCV)E1 protein.Methods The human pancreas cDNA library was amplified,purified and evaluated,and then the purified library plasmids were transformed into yeast strain Y187.The reconstructed plasmid pGBKT7-E1 was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp.The transformed AH109 mated with Y187 that contained the library plasmids.The diploid yeast cells were plated on nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-α-gal for selecting.The plasmids in diploid yeast cells were extracted and electrotransformed into E.coli DH5α.The plasmids in DH5α were extracted,sequenced and blasted.Result Sixteen proteins interacting with HCV E1 were found.Conclusion Some of the sixteen pancreatic proteins may be related with metabolisms of glucose and lipid.
5.Clinical diagnosis and treatment of renal clear cell carcinoma combined with papillary renal cell carcinoma in unilalateral kidney
Jianshen CHEN ; Ming LI ; Yueqing WU ; Mao TANG ; Jinqian LI ; Xinwu GUO
International Journal of Surgery 2018;45(5):314-318
Objective To investigate the clinical imaging features and immune manifestations of clear cell carcinoma and papillary renal cell carcinoma at the same time in one side of the kidney,in order to increase the awareness of the disease and improve the clinical diagnosis and treatment.Methods This study retrospectively analyzed the clinical data of 7 patients with unilateral renal clear cell carcinoma combined with papillary cell carcinoma admitted to the Department of Urology,People's Hospital of Xinjiang Uygur Autonomous Region from January 2007 to June 2017.The general data,images performance,surgical methods,immunohistochemical results of the patients were observed.Results In 7 patients with multiple tumors of ipsilateral kidney,6 were males,accounting for 85.71% (6/7);1 female,accounting for 14.29% (1/7).Ages 45-78 years old,average 63 years old;4 cases were found by physical examination,accounting for 57.14% (4/7);2 cases were admitted to hospital with obvious oncological symptoms,accounting for 28.57% (2/7);1 case of renal nonfunction,accounting for 14.29% (1/7).The tumor was located in 6 cases of left kidney,accounting for 85.71% (6/7),and the tumor was located in 1 case of right kidney,accounting for 14.29% (1/7).Preoperative imaging examination clearly indicated multiple tumors in 3 cases,and the remaining 4 cases only showed placeholders.Lesions,of which the maximum diameter of the tumor 7.5 cm,2 patients with a diameter of > 4 cm;4 cases of radical renal resection and partial nephrectomy;3 cases of clear cell carcinoma Fuhrman Ⅰ grade,Fuhrman Ⅱ grade 5 cases postoperative pathology report There were 4 cases of papillary renal cell carcinoma in Fuhrman grade Ⅰ and 3 cases in Fuhrman grade Ⅱ,all of them were type Ⅰ cell carcinoma.Conclusions Ipsilateral kidney associated with papillary renal cell carcinoma and clear cell carcinoma is rare clinically,its formation is unknown.Readings should be repeated before surgery.If you can identify multiple tumors,you will be able to choose the surgical procedure.Partial nephrectomy or radical nephrectomy may be performed depending on the size of the tumor.Radical nephrectomy is recommended.
6.The change in IL-13 mRNA expression in rat lungs with acute pulmonary injury induced by lipopolysaccharide.
Qi LI ; Guisheng QIAN ; Qing ZHANG ; Jianyue XU ; Yong LONG ; Zhengcai TANG ; Jinqian GONG
Chinese Journal of Burns 2002;18(3):145-148
OBJECTIVETo observe the change in IL-13 mRNA expression in rat lungs with acute lung injury (ALI) induced by lipopolysaccharide (LPS), so as to investigate the significance of anti-inflammatory mechanism in the development of ALI.
METHODS(1) Different doses (2 approximately 8 mg/kg) of LPS were injected via the tail vein to the rats to establish ALI model. (2) The IL-13 mRNA content in pulmonary tissue was determined by RT-PCR.
RESULTS(1) Different degrees of ALI was induced in the rats by different doses of LPS. ARDS was produced in the rats by LPS of 6 mg/kg. (2) The IL-13 mRNA expression in the pulmonary tissue could be enhanced by the induction of LPS, especially when its dose was larger than 6 mg/kg.
CONCLUSION(1) Rat ALI model could be produced by LPS injection via the tail vein. (2) LPS in dose of 6 mg/kg was the threshold for the production of rat ARDS. (3) The incidence of rat ARDS might be closely related to the enhanced expression of IL-13 mRNA in rat pulmonary tissue.
Acute Disease ; Animals ; Disease Models, Animal ; Female ; Interleukin-13 ; biosynthesis ; genetics ; Lipopolysaccharides ; Male ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Respiratory Function Tests ; Respiratory Insufficiency ; chemically induced ; metabolism ; physiopathology
7.Anesthesia for cesarean section after failed labor under combined spinal and epidural analgesia: analysis of 58 cases
Jinqian HU ; Ailin LUO ; Li WAN ; Dongji HAN
Chinese Journal of Perinatal Medicine 2019;22(2):123-126
Objective To summarize the experience in anesthesia management for cesarean section(CS) after failed labor in women with combined spinal and epidural analgesia and to provide evidence for more effective and safer clinical practice.Methods We included 58 singleton pregnant women who received spinal and epidural analgesia during labor but converted to CS at Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from October 2017 to October 2018.When CS was indicated,10 ml of 0.125% ropivacaine was given through the epidural catheter immediately,and 5 ml of 1% lidocaine was given when arriving at the operating room.Five minutes later,10 ml of 1% ropivacaine was administered.General anesthesia would be required when epidural anesthesia was considered to be failed 10 min after the usage of ropivacaine.Clinical datas were retrospectively reviewed.The management and effects of anesthesia for CS,maternal and neonatal outcomes were described.Results Of the 58 patients when CS was indicated during labor,5.2%(3/58) received general anesthesia immediately.Among the other 94.8% (55/58) who received epidural anesthesia,3.6%(2/55) converted to general anesthesia later.Adverse effects of epidural anesthesia included reduced mean artery pressure (8/53,15.1%),nausea and vomiting (3/53,5.7%).For the neonatal Apgar score at 1 min,one out of the 58 babies (1.7%) was between 0 and 3,ten (17.2%) between 4 and 7,and 47 (81.0%) between 8 and 10.The two babies (3.4%) with Apgar score between 4 and 7 at 5 min were both referred to the department of Neonatology,and the rest 56 neonates (96.6%) scored 8-10.Conclusions Spinal and epidural analgesia in labor can be safely and effectively adjusted to anesthesia for CS.
8.Clinical effect of iliac artery interventional chemotherapy combination with bladder perfusion for high level of bladder urothelial carcinoma
Jiansen CHEN ; Ming LI ; Yueqing WU ; Mao TANG ; Zetian TANG ; Xinwu GUO ; Jinqian LI ; Kesong QIN
International Journal of Surgery 2017;44(9):583-586,后插1
Objective To evaluate the clinical effect of iliac artery interventional chemotherapy combination with bladder perfusion for high level of bladder urothelial carcinoma after transurethral bladder tumor electricity cut operation for those who intend to retain the bladder.Methods From February 2010 to December 2016,a total of 74 cases high-grade urothelial carcinoma of the bladder,giving chemotherapy and suitted support were retrospectively analyzed.The patients were divided into two groups according to different chemotherapy methods.Artery perfusion chemotherapy group:43 cases treated by transurethral bladder tumor cutting + bilateral iliac artery interventional perfusion chemotherapy + bladder perfusion in combination therapy;Intravenous chemotherapy group:31 cases underwent transurethral bladder tumor cut method + cisplatin,vein gemcitabine + bladder perfusion chemotherapy combined therapy.According to the ((x) ± s) between the groups.The method of count data is represented by chi-square test.Kaplan-meier method was used to describe the survival time of two groups of patients after operation.Results Two groups of patients with postoperative there was no statistically significant difference in disease-free survival time.In the bone marrow suppression (x2 =4.956,P =0.029);gastrointestinal tract reaction (x2 =5.912,P =0.012);dermatitis,mucositis (x2 =4.276,P =0.013),etc.,has the advantages in the iliac artery interventional perfusion chemotherapy (P < 0.05).Conclusion Via efficient iliac arterial infusion chemotherapy in patients with venous chemotherapy patients,to the retention of the bladder has a certain value,and the incidence of adverse reactions after surgery is low,have certain advantages.
9.Perioperative factors related to prognosis of regular hepatectomy in comparison with irregular hepatectomy.
Zhiqiang FENG ; Hongqi LI ; Jinqian ZHANG ; Lining XU ; Mei XIAO ; Zhiqiang HUANG ; Hongyi ZHANG
Chinese Medical Journal 2014;127(2):239-245
BACKGROUNDThe aim of this research was to analyze the perioperative factors of regular hepatectomy and irregular hepatectomy. The superiority of the clinical application of the two methods was compared in the perioperative period.
METHODSFrom 1986 to 2011, 1798 patients underwent consecutive liver resections with regular hepatectomy and irregular hepatectomy at the Air Force General Hospital of People's Liberation Army and the General Hospital of Chinese People's Liberation Army. Their medical documentation was investigated retrospectively.
RESULTSIn patients on whom regular hepatectomy and irregular hepatectomy were performed, there was no significant difference in perioperative blood loss, complications, in-hospital mortality, hospital stay, and so on. But in regular hepatectomy, operating time was an independent risk factor (P < 0.001, OR = 1.004).
CONCLUSIONSThere was no significant difference between the perioperative risk of regular hepatectomy and that of irregular hepatectomy.
Female ; Hepatectomy ; adverse effects ; methods ; Humans ; Male ; Perioperative Period ; Retrospective Studies
10.Targeting cAMP in D1-MSNs in the nucleus accumbens, a new rapid antidepressant strategy.
Yue ZHANG ; Jingwen GAO ; Na LI ; Peng XU ; Shimeng QU ; Jinqian CHENG ; Mingrui WANG ; Xueru LI ; Yaheng SONG ; Fan XIAO ; Xinyu YANG ; Jihong LIU ; Hao HONG ; Ronghao MU ; Xiaotian LI ; Youmei WANG ; Hui XU ; Yuan XIE ; Tianming GAO ; Guangji WANG ; Jiye AA
Acta Pharmaceutica Sinica B 2024;14(2):667-681
Studies have suggested that the nucleus accumbens (NAc) is implicated in the pathophysiology of major depression; however, the regulatory strategy that targets the NAc to achieve an exclusive and outstanding anti-depression benefit has not been elucidated. Here, we identified a specific reduction of cyclic adenosine monophosphate (cAMP) in the subset of dopamine D1 receptor medium spiny neurons (D1-MSNs) in the NAc that promoted stress susceptibility, while the stimulation of cAMP production in NAc D1-MSNs efficiently rescued depression-like behaviors. Ketamine treatment enhanced cAMP both in D1-MSNs and dopamine D2 receptor medium spiny neurons (D2-MSNs) of depressed mice, however, the rapid antidepressant effect of ketamine solely depended on elevating cAMP in NAc D1-MSNs. We discovered that a higher dose of crocin markedly increased cAMP in the NAc and consistently relieved depression 24 h after oral administration, but not a lower dose. The fast onset property of crocin was verified through multicenter studies. Moreover, crocin specifically targeted at D1-MSN cAMP signaling in the NAc to relieve depression and had no effect on D2-MSN. These findings characterize a new strategy to achieve an exclusive and outstanding anti-depression benefit by elevating cAMP in D1-MSNs in the NAc, and provide a potential rapid antidepressant drug candidate, crocin.